We propose to carry out studies of the operation and control of the purine nucleotide cycle in perfused skeletal muscle. Fifty preliminary perfusion experiments show that our preparations exhibit normal metabolic parameters. At rest they produce little ammonia. Electrical stimulation results in a considerable output of ammonia. The objective is to understand the conditions under which the purine nucleotide cycle operates, and how its operation affects the control of glycolysis and the citric acid cycle. We propose to study how the interactions of the purine nucleotide cycle with glycolysis lead to oscillations in both pathways. The effect of variables such as pH and the degree of energy drain on the rates of operation of the purine nucleotide cycle and glycolysis, and on the type of oscillation, will be investigated in extracts prepared from different types of muscles, in different nutritional and hormonal states. Studies demonstrating the operation of the cycle in extracts of brain and kidney will be completed. We will determine whether the purine nucleotide cycle can operate at a rate adequate to account for the maximum rates of ammonia production by brain and kidney. (Preliminary measurements indicate it can.) The control properties of the cycle in brain extracts will be studied because they probably differ from those of muscle. The regulatory properties of adenylosuccinate synthetase and adenylate deaminase will be studied further so that the factors that control the operation of the purine nucleotide cycle can be better understood. BIBLIOGRAPHIC REFERENCES: "Purine Nucleotide Cycle. VI. Evidence for the Occurrence of the Cycle in Brain" by Vera Schultz and John M. Lowenstein, J. Biol. Chem. (1976) 250, Jan. or Feb. "The Purine Nucleotide Cycle. VII. A Pathway for Ammonia Production in the Rat Kidney" by Ronald T. Bogusky, Leah M. Lowenstein, and John M. Lowenstein, Journal of Clinical Investigation (1976), submitted for publication.